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Fig. 3. Axin2 deficiency stimulates expansion of osteoprogenitors. The
osteoblast precursors undergoing active divisions in the
Axin2+/+ (A) and Axin2–/–
suture (B) were identified by the expression of Ki67. Immunohistochemical
staining reveals increases in the presence of the mitotic osteoprogenitors in
the Axin2–/– suture at P8. In the defined
suture region (broken lines), osteoprogenitors positive (brown) and negative
(blue) for Ki67 were counted to measure proliferation abnormalities. At least
six different regions from the same developmental stage were analyzed to
obtain the average percentage of Ki67 positive (Ki67+) cells at E16.5, P8 and
P17 (C). Primary osteoblast precursors isolated from nasal and frontal bones
of the Axin2+/+ (D) and
Axin2–/– (E) were grown in culture media for
in vitro BrdU incorporation analysis. Cells were immunostained with
-BrdU antibody (brown), and counterstained with Hematoxylin (blue). The
stained images were taken randomly to determine the percentage of
proliferating cells by counting the BrdU-positive cells in a total of 1000
cells. The graph shows the average percentage of Axin2+/+
and Axin2–/– in three independent experiments
(F). Scale bars: 50 µm in A,B; 200 µm in D,E.
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